1. Field of the Invention
The present invention relates to a method for purifying a protein, particularly enzymes from biomass, particularly the enzyme penicillin amidase.
2. Description of the Background
It is known that proteins, particularly enzymes, can be purified by way of hydrophobic chromatography. For example, the enzyme penicillin amidase (penicillin acylase PcA) can be purified from E. coli (E.C. 3.5.1.11), which is used for technical conversion of fermentatively produced penicillins in 6-amino penicillin acid (df. H. J. Rehm & G. Reed in Biotechnology, Vol. 7a, p. 168, 169 VCH 1987), on phenyl glycine sepharose, by means of affinity chromatography (Indian Patent 15 50 51A, Purification of Penicillin Acylase Enzyme, P. S. Borkar and P. B. Majahan). Furthermore, benzyl EUPERGIT is used for purification of PCA by affinity chromatography. Affinity chromatography with affinity ligands consisting of phenylacetic acid or its derivatives is described in Chem. Abstracts 97, 35363a. Other methodology employs conjugates of agarose or sepharose with ampicillin or amoxicillin as matrices for affinity chromatography (Indian Patent 155 050; P.B. Mahajan et al., Hind. Antibiot. Bull. 24 (1-2) 38, 1982, Chem. Abstracts97, 87551e as well as Chem. Abstracts95, 110 740 K).
The methods which have been proposed thus far are complicated, without exception, both with regard to the number of individual operations required and with regard to the apparatus necessary to perform the operations. In fact the significant time required for protein purification is unsatisfactory. A need therefore continues to exist for a simplified method of protein and purification isolation which is fast and involves minimal loss of protein.